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Free Content Polyclonal expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract

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Human immunodeficiency virus (HIV) -specific T-cell responses are detectable in the female genital tract of HIV-infected women but little is known about their frequency or the factors that influence their detection. We investigated the feasibility of polyclonal in vitro expansion of cervical cytobrush-derived T cells to investigate HIV-specific responses in the female genital tract in HIV-infected women. Cytobrush-derived cervical cells were isolated from 22 HIV-infected women and expanded with anti-CD3 and recombinant interleukin-2. Cervical T-cell lines were investigated for Gag-specific responses by interferon-γ ELISPOT and compared with those detected in matched blood samples. Cervical T-cell lines were established from 16/22 (72·7%) participants. Although the absolute number of CD3± cells recovered after expansion was positively associated with the number of cells isolated ex vivo (P = 0·01; R = 0·62), we observed a significant negative correlation between fold expansion and ex vivo cell number (P = 0·004; R = −0·68). We show that both the magnitude (P = 0·002; R = 0·7) and specific Gag regions targeted by cervical T-cell lines (P < 0·0001; R = 0·5) correlated significantly with those detected in blood. With one exception, cervical interferon-γ T-cell responses to Gag were detected only in HIV-infected women with blood Gag-specific response > 1000 spot-forming units/106 cells. We conclude that cervical Gag-specific T-cell responses in expanded lines are most easily detectable in women who have corresponding high-magnitude Gag-specific T-cell responses in blood.
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Keywords: T cell; cervical; expansion; gag; human immunodeficiency virus; mucosal

Document Type: Research Article

Affiliations: 1: Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town 2: Department Obstetrics and Gynaecology, University of Cape Town and Groote Schuur Hospital, Observatory, Cape Town

Publication date: May 1, 2010

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